OBJECTIVETo optimize the methods of isolating human eccrine sweat gland cells in vitro so as to get efficiently primary human sweat glands.

METHODSThe fresh and normal skin tissue was cut into pieces of microskin about 1mm3 and the following 3 group digestion buffer was applied to isolated gland cells. The digestion buffer of group A was the equivoluminal mixture of Trypsin-Ethylene Diamine Tetraacetic Acid (EDTA) and collagenase-II (2 mg/ml). The digestion buffer of group B was collagenase-II (2 mg/ml) traditionally and group C was Trypsin-EDTA. These three groups were placed into an incubator simultaneously and the emerging time of dissociated sweat glands was calculated. Sweat glands were sorted out and then placed in culture dish. The adherence and the growth of cells were observed. The proliferation index was detected by flow cytometry. The identification of cultured cells was performed by immunocytochemical staining.

RESULTSAfter digesting 30 min in group A and C, a very few of dissociated sweat glands were emerging. But after digesting for 2 h, there were lots of dissociated sweat glands emerging in group A rather than in group C. The emergence of dissociated sweat glands in group B would require at least 6 hours. After seeded in culture dishes, the sweat glands in group C couldn't adhere to the wall of dish, but the sweat glands in group A and B adhered very well and even grew like paving stones after 9 days. In addition, the proliferation index were (18 ± 4) % and (17 ± 6) % respectively, there was no statistical difference. The results of immunocytochemical staining showed that the cells expressed carcino-embryonic antigen (CEA) and cytokeratin 7(CK7) in group A and B.

CONCLUSIONTrypsin-EDTA combined with collagenase-II can shorten the time of isolating sweat gland cells and have no effect on cell activity and proliferation.

Cell Separation ; methods ; Cells, Cultured ; Eccrine Glands ; cytology ; Humans ; In Vitro Techniques

OBJECTIVETo study the anticancerous effect of Fuganchun 6 (FGC-6) and its immunoregulatory effect on tumor-bearing mice.

METHODThe mice inoculated by H22 cells were divided into 5 groups: model group, 5-Fu group and FGC-6 in high dose, medium dose, and low dose groups. The normal mice were also observed. These mice were treated for 10 days. The weight of tumor mass and mouse were examined. The target-cell-killing activity of NK cells. The proliferation activity of lymphocyte and the production of IL-2 of murine splenocytes were detected respectively. The serum containing FGC-6 was prepared and its inhibition effect on H22 cells was examined by MTT assay and growth curve in vitro.

RESULTGrowth of tumor was inhibited markedly by FGC-6 high dose. The inhibition of serum containing FGC-6 on the proliferation of H22 cells in vitro was observerd in a dose and time-dependent manner. The target-cell-killing activity of NK cells and the production of IL-2 of murine splenocytes of model group were lower than those of normal group (P < 0.05). When compared with model group, FGC-6 in high dose elevated the two indexes above-mentioned, and also enhanced the proliferation activity of lymphocyte markedly (P < 0.05). The production of IL-2 of murine splenocytes was also improved when treated by FGC-6 in medium dose (P < 0.05).

CONCLUSIONFGC-6 can inhibite the growth of H22 cells markedly and also can strengthen the immunity of H22 transplanted mouse.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Cell Line ; Cell Proliferation ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Interleukin-2 ; metabolism ; Killer Cells, Natural ; immunology ; Liver Neoplasms, Experimental ; immunology ; pathology ; Lymphocytes ; pathology ; Male ; Mice ; Plants, Medicinal ; chemistry ; Spleen ; cytology ; metabolism

Objective To study the effect of family intervention on heart rate variability in patients with coronary heart disease (CHD).Methods A total of 92 CHD patients were selected according to the enrollment criteria.They were randomly divided into two groups, intervention group with 46 patients, and control group with 46 patients.The intervention group accepted family intervention while the control group accepted only general health education.The heart rate power spectral density and time-domain were analyzed with 24 hour electrocardiographic monitoring.Results The heart rate variability was improved in both groups (intervention group P <0.01, control group P <0.05).Compared with the control group,time-domain analysis in intervention group revealed a significant improved in SDNN, SDANNI, SDNNI, RMSSD, PNN50 ( P < 0.05 ).Frequency domain analysis revealed marked improve of VLE、LF、HF (P <0.05).Conclusions Family intervention can significantly improve the heart rate variability in CHD.

Objective To explore the feasibility for modeling and forecasting outpatient consulta-tionrate of influenza-like illness ( ILI ) in Huangpu District of Shanghai . Methods ILI consultation rates from the first week of 2011 to the 52 th week of 2014 were collected through the National Influenza Surveillance Information System .SAS 9.3 was used to establish an optimal ARIMA model of ILI data .The forecasting ability of the ARIMA model was afterwards evaluated by using data from the first to the sixth week in 2015 . Results ARIMA ( 1 ,0 ,0 ) was identified as the final model in fitting outpatient vising rate of ILI with stable and white noise residual .Model validation showed that the real outpatient-visiting rate of ILI all fell in the 95%confidence interval of the predictive values . Conclusion ARIMA (1,0, 0 ) could be used in the forecast of ILI consultation rates in Huangpu District of Shanghai .

Most patients with differentiated thyroid cancer have a good prognosis after radioiodine-131 therapy,but a small number of patients are insensitive to radioiodine-131 therapy and even continue to develop disease.At present,some targeted drugs can improve progression-free survival in patients with radioactive iodine-refractory differentiated thyroid cancer(RAIR-DTC),such as sorafenib and levatinib,have been approved for the treatment of RAIR-DTC.However,due to the presence of primary and acquired drug resistance,drug efficacy in these patients is unsatisfactory.This review introduces the acquired drug resistance mechanism of sorafenib in the regulation of mitogen-activated protein kinase(MAPK)and phosphatidylinositol-3-kinase(PI3K)pathways and proposes related treatment strategies,in order to provide a reference for similar drug resistance mechanism of sorafenib and effective treatment of RAIR-DTC.

Objective To understand the epidemiological characteristics of COVID-19 epidemic in Huangpu District of Shanghai, and to provide scientific evidence for prevention and control of COVID-19. Methods Descriptive statistics were used to study the suspected and confirmed cases of COVID-19 reported from January 21 through March 10, 2020 in Huangpu District, Shanghai. Results A total of 120 suspected cases of COVID-19 were reported, of which 12 were diagnosed and 108 were excluded.The first confirmed case was reported on January 21, and the last case was on February 10; the majority (11/12) of the confirmed cases were reported from January 21 through February 1.The average duration of time from the symptom onset to the first medical visit was 2.6 days, whereas the average duration from the first medical visit to the hospital diagnosis was 2.2 days.There were 15 suspected cases with a confirmed history of residence or tourism in Wuhan, in which 6 were confirmed cases.Moreover, 5 suspected cases had a confirmed history of contact with other confirmed cases, in which 3 were confirmed cases.Thus, exposure in Wuhan and exposure to confirmed cases were the most significant risk factors at this stage of the epidemic. Conclusion The 12 cases identified in Huangpu District of Shanghai are all adults, half of whom had confirmed history of exposure in Wuhan.The first cluster of COVID-19 cases in Shanghai is documented in Huangpu District.Epidemiological investigation reveals that the confirmed cases might be infectious the day before the symptom onset.

BACKGROUNDTo study arboviruses carried by mosquitoes in Liaoning Province in 2002.

METHODSTotally 4927 mosquitoes were collected from Liaoning Province in July 2002. Virus strains were isolated by inoculating BHK-21, C6/36 and Vero cells. The newly isolated strains were identified by serological (ELISA and IFA) and molecular methods (Real-Time PCR and RT-PCR).

RESULTSTwo strains were isolated from mosquitoes causing cytopathogenic effect (CPE) on cells and were fatal to suckling mice. Serological tests showed that both were positive for the antibody to JEV. The PrM and E gene were cloned and sequenced. The phylogenetic analysis showed that the new isolates belonged to genotype I, JEV. Sequence analysis showed that the homology of nucleotide sequences and amino acid (AA) sequences between the two strains was 100%. Compared with the nucleotide sequences between the two strains and the standard JEV vaccine strain SA14-14-2, the difference was up to 4.11%, and the difference of AA was 0.6%.

CONCLUSIONTwo strains of JEV were isolated and identified in Liaoning province, both belonged to genotype I JEV.

Animals ; Cell Line ; Cercopithecus aethiops ; China ; Culicidae ; virology ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Analysis, DNA ; Vero Cells ; Viral Envelope Proteins ; genetics

OBJECTIVETo investigate the therapeutic effectiveness of intracoronary implantation of autologous bone marrow mononuclear cells (BM-MNC) in miniswine model of reperfused myocardial infarction.

METHODSSixteen miniswine myocardial ischemic reperfusion injury models made by ligation of the distal one third segment of left anterior descending artery for 90 minutes were randomized into 2 groups. In BM-MNC group (n = 9), (3.54 +/- 0.90) X 10(8) BM-MNC were intracoronary injected, and in the control group (n = 7), phosphate buffered saline was injected by the same way. Echocardiographic and hemodynamic results, vessel density, and myocardial infarction size were evaluated and compared before and 4 weeks after cell transplantation.

RESULTSIn BM-MNC group, there were no differences between before and 4 weeks after transplantation in aspects of left ventricular ejection fraction (LVEF), interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, or +dp/dtmax. In control group, LVEF, interventricular septal thickness, left ventricular lateral and anterior septal wall thickness, cardiac output, and +dp/dtmax decreased significantly 4 weeks after transplantation (P < 0.05). Left ventricular end-diastolic pressure and -dp/dtmax, did not change significantly before and after cell transplantation in both groups. Capillary density in BM-MNC group was greater than that in control group [(13.39 +/- 6.96)/high power field vs. (3.50 +/- 1.90)/high power field, P < 0.05]. Infarction area assessed by tetrazolium red staining and the infarction percentage decreased in BM-MNC group compared with those in control group (P < 0.05).

CONCLUSIONSTransplantation of BM-MNC into myocardium with ischemic reperfusion injury increases capillary density and decreases infarction area. It has significantly beneficial effect on cardiac systolic function rather than on diastolic function.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Marrow Transplantation ; Capillaries ; physiology ; Echocardiography ; Heart ; anatomy & histology ; physiology ; physiopathology ; Hemodynamics ; Myocardial Ischemia ; Random Allocation ; Swine ; Systole ; physiology ; Transplantation, Autologous ; physiology

OBJECTIVETo simulate and assess the clinical effect of intracoronary infusion of bone marrow mononuclear cells or peripheral endothelial progenitor cells on myocardial reperfusion injury in mini-swine model.

METHODSTwenty-three mini-swine with myocardial reperfusion injury were used as designed in the study protocol. About (3.54 +/- 0.90) x 10(8) bone marrow mononuclear cells (MNC group, n = 9) or (1.16 +/- 1.07) x 10(7) endothelial progenitor cells (EPC group, n = 7) was infused into the affected coronary segment of the swine. The other mini-swine were infused with phosphate buffered saline as control (n = 7). Echocardiography and hemodynamic studies were performed before and 4 weeks after cell infusion. Myocardium infarction size was calculated. Stem cell differentiation was analyzed under a transmission electromicroscope.

RESULTSLeft ventricular ejection fraction dropped by 0% in EPC group, 2% in MNC group, and 10% in the control group 4 weeks after cell infusion, respectively (P < 0.05). The systolic parameters increased in MNC and EPC groups but decreased in the control group. However, the diastolic parameters demonstrated no significant change in the three groups (P > 0.05). EPC decreased total infarction size more than MNC did (1.60 +/- 0.26 cm2 vs. 3.71 +/- 1.38 cm2, P < 0.05). Undermature endothelial cells and myocytes were found under transmission electromicroscope.

CONCLUSIONSTransplantation of either MNC or EPC may be beneficial to cardiac systolic function, but might not has obvious effect on diastolic function. Intracoronary infusion of EPC might be better than MNC in controlling infarction size. Both MNC and EPC may stimulate angiogenesis, inhibit fibrogenesis, and differentiate into myocardial cells.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Cell Differentiation ; Endothelial Cells ; cytology ; Myocardial Reperfusion Injury ; pathology ; therapy ; Myocardium ; pathology ; Stem Cells ; cytology ; Swine ; Swine, Miniature

Objectives: This study aimed to observe the change of arachidonic acid-induced platelet aggregation rate (AA-Ag) and short-term adverse reactions after taking 50 or 100 mg/d aspirin(enteric-coated sustained-release formulation) or 100 mg/d aspirin (enteric-coated aspirin tablet)in the elderly Chinese population (aged 60 years or older). Methods: A total of 1 194 participants aged 60 or older, who should be recommended to take aspirin therapy due to medical reasons, were recruited and randomly assigned into three groups to receive enteric-coated sustained-release aspirin tablet (50 mg, once daily, group A), or 100 mg, once daily (group B) or enteric-coated aspirin tablet 100 mg once daily (group C), respectively. AA-Ag was measured after (14±3)days of aspirin treatment. Adverse events and bleeding events were recorded during the (28±3)days of follow-up. Results: The AA-Ag in group A (n=347), B (n=338) and C (n=332) post 14-day aspirin therapy were 6.65 (4.03,10.84)%, 5.89(3.22,10.03) % and 6.00(3.68,10.09) %, respectively (P>0.05). During the 28 days follow-up, the adverse events rate of group A (n=388), B (n=387) and C (n=385) was 3.87%,3.36%, and 7.95%, and the mild bleeding events rate was 3.09%, 2.33%, and 6.23%, respectively. Adverse events rate and mild bleeding events rate were significantly higher in group C than in group A and B (P<0.05). Conclusions: Compared with 100 mg-dose aspirin, 50 mg-dose aspirin achieves similar anti-platelet aggregation effect in this elderly Chinese population. The short-term adverse events and mild bleeding risk of aspirin with enteric-coated sustained-release formulation were fewer than that of enteric-coated formulation.